1.1 本试验方法规定了对细菌生物膜进行定量液体消毒剂功效试验所需的操作参数。 1.2 对试验方法进行了优化和验证 铜绿假单胞菌 或 金黄色葡萄球菌 CDC生物膜反应器中生长的生物膜( E3161 ). 该方法适用于评估使用具有类似试片尺寸的方法（如实践）中概述的程序生长的其他细菌 E3161 ，试验方法 E2562 ，或试验方法 E2196年 . 1.3 根据制造商的使用说明，在评估中使用消毒剂制备和接触时间。 1.4 该试验方法使用封闭系统处理生物膜。 将试片放置在单管中，用于处理、中和和收获步骤，以防止细胞损失。 1.5 本试验方法描述了一种采集和分析程序，包括涡流和超声波处理和未处理的对照生物膜，以及使用过滤来回收可培养细胞，以降低检测限。生物膜种群密度记录为对数 10 每片菌落形成单位。疗效以对数形式报告 10 可培养细胞的减少。 1.6 进行该分析需要基本微生物学培训。 1.7 以国际单位制表示的数值应视为标准值。本标准不包括其他计量单位。 1.8 本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.9 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 单管法旨在按照实践中概述的程序评估消毒剂对CDC生物膜反应器中生长的生物膜的效果 E3161 . CDC反应器中生长的生物膜是有利于生物膜形成的表面在高流体剪切下形成的生物膜的代表。 5.1.1 营养生物膜细菌在表型上不同于相同基因型的悬浮浮游细胞。生物膜生长反应器旨在产生具有特定特性的生物膜 ( 2. ) . 改变工程系统或操作条件将改变这些特性以及物理化学环境。生物膜研究和测试的目标是为特定研究选择生成最相关生物膜的生长反应器和操作条件。 5.2 该测试方法旨在确定对数 10 在封闭系统中暴露于消毒剂后的细菌减少。 5.3 试验方法使用50毫升锥形管。锥形几何形状允许消毒剂暴露在试片所有表面的生物膜中。对于发泡消毒剂或需要更大体积中和剂的消毒剂，使用250 mL锥形管，其保持所需的几何形状并允许更大的中和能力。 5.4 每个试验包括三个未经处理的对照试片（暴露于缓冲稀释水）和五个经处理的试片（每个消毒剂/浓度/接触时间组合）。

1.1 This test method specifies the operational parameters required to perform a quantitative liquid disinfectant efficacy test against bacterial biofilm. 1.2 The test method was optimized and validated for a Pseudomonas aeruginosa or Staphylococcus aureus biofilm grown in the CDC Biofilm Reactor ( E3161 ). The method is suitable for evaluating additional bacteria grown using the procedures outlined in methods with comparable coupon dimensions such as Practice E3161 , Test Method E2562 , or Test Method E2196 . 1.3 Disinfectant preparation and contact time are used in the assessment according to the manufacturer’s instructions for use. 1.4 The test method uses a closed system to treat biofilm. A coupon is placed in a single tube for the treatment, neutralization, and harvesting steps to prevent the loss of cells. 1.5 This test method describes a harvesting and analysis procedure which includes vortexing and sonicating treated and untreated control biofilm, and recovery of culturable cells using filtration to lower the limit of detection. Biofilm population density is recorded as log 10 colony-forming units per coupon. Efficacy is reported as a log 10 reduction of culturable cells. 1.6 Basic microbiology training is required to perform this assay. 1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. 1.9 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 5.1 The Single Tube Method is designed to evaluate the efficacy of disinfectants against biofilm grown in the CDC biofilm reactor following the procedures outlined in Practice E3161 . Biofilm grown in the CDC reactor is representative of biofilm that forms under high fluid shear on surfaces conducive to biofilm formation. 5.1.1 Vegetative biofilm bacteria are phenotypically different from suspended planktonic cells of the same genotype. Biofilm growth reactors are engineered to produce biofilm with specific characteristics ( 2 ) . Altering either the engineered system or operating conditions will modify those characteristics as well as the physicochemical environment. The goal in biofilm research and testing is to choose the growth reactor and operating conditions that generate the most relevant biofilm for the particular study. 5.2 The test method was designed to determine the log 10 reduction in bacteria after exposure to a disinfectant in a closed system. 5.3 The test method uses 50 mL conical tubes. The conical geometry allows for disinfectant exposure to biofilm on all surfaces of the coupon. For foaming disinfectants or for disinfectants requiring a larger volume of neutralizer, 250 mL conical tubes are used which preserve the required geometry and allow for greater neutralization capacity. 5.4 Each test includes three untreated control coupons (exposed to buffered dilution water) and five treated coupons (per disinfectant/concentration/contact time combination).